WO2000002574A1 - Powdery preparation for mucosal administration containing polymeric medicine - Google Patents

Abstract

A powdery preparation for mucosal administration which comprises a polymeric medicine and a cationic polymer. The preparation is formed by adding a cationic polymer [especially an aminoalkyl methacrylate copolymer or poly(vinyl acetal diethylaminoacetate)] to a polymeric medicine, optionally further adding a thickening polymer, and powdering the mixture. Thus, the polymeric medicine can be effectively absorbed through a mucous membrane.

Description

 Description Powdered transmucosal formulation containing polymer drug Technical field

 The present invention relates to a transmucosal administration preparation containing a polymer drug as an active ingredient, and more particularly, to a powdery transmucosal administration preparation containing a polymer drug and a cationic polymer. The present invention particularly relates to a powdery nasal mucosal administration preparation. Background art

 At present, high-molecular-weight drugs are administered intravenously or subcutaneously by injection during treatment. However, administration of macromolecular drugs by injection is difficult and painful for the patient himself, so transmucosal administration is desired as a simpler administration method than injection. Specific examples of transmucosal administration include administration from the nasal mucosa, ocular mucosa, oral mucosa, lung mucosa, vaginal mucosa, and gastrointestinal mucosa such as gastric mucosa, small intestinal mucosa, large intestinal mucosa, and rectal mucosa. Can be. In particular, nasal mucosal administration has attracted attention as a relatively simple administration method that can provide rapid drug absorption and a reliable effect. However, its absorbency depends on the molecular weight, and those with a molecular weight of less than 1,000 are absorbed relatively effectively, but above that, it is difficult to absorb effectively without any contrivance. (C.McMartin et al .: J.Pharm.Sci., 76 (7): 535-540 (1968)). Therefore, it was difficult to obtain a therapeutic effect by administering a polymer drug through the nasal mucosa.

As a means of improving the poor absorbability of macromolecular drugs, a method using a combination of surfactants and bile salts as absorption enhancers (S. Hirai et al .: Int. J. Pharma., 9 : 1 7 3-18 4 (1 9 8 1), Y. M aitani Etc .: Drug Design and Remove, 1: 65-70 (1966)), a method using cyclodextrin in combination as an absorption promoter (NGSM chipper, etc .: J Control R elease, 2 1 (1): 17 3-18 5 (1992), T. I rie, etc.: J. Inter. Pharm., 84: 1229-1. 39 (1992)) There is a problem that these absorption enhancers may cause damage to the nasal mucosa. On the other hand, a method in which polymers such as albumin, dextran, and sodium hyaluronate are used in combination as absorption enhancers (T. Igawa et al .: Chem. Pharma. Bull. 36 (8 ): 305-5-359 (1998), JP-A-6-65090, JP-A-8-198772), but the effect of promoting absorption is still lacking. Practical use has not yet been achieved due to inadequacy and difficulty in industrial production.

 In Japanese Patent Application Laid-Open No. 10-95738, fluorescein isothiosine-to-dextran (hereinafter referred to as “FITC-dextran”, molecular weight: 4,400) is used as a poorly absorbable model drug (substance). A preparation was prepared by adding this to physiological saline in which arginine, its poly-form, or a salt of the poly-form was dissolved, and this formulation was administered into the nasal mucosa of Wistarrat. It is stated that high blood levels persisted.

 JP-T-Hei 4-503508 describes that a formulation containing DEAE-dextran or chitosan added to an insulin solution was administered to the nostrils of rats.

 Although various methods as described above have been developed, there is still a need for more effective and practical methods as a means for improving the poor absorbability of polymer drugs.

Accordingly, an object of the present invention is to provide a transmucosal administration preparation, particularly a nasal administration preparation, which can safely and efficiently absorb a polymer drug from mucosa. Another object of the present invention is to provide a powdery pharmaceutical composition capable of safely and efficiently absorbing a polymer drug into a living body. Disclosure of the invention

 The present inventors have studied for the purpose of developing a formulation for safely and efficiently absorbing a high-molecular-weight drug through the mucous membrane.As a result, the cationic polymer expands the tight junction of the mucosal tissue by increasing the tight junction. Promote the absorption of pharmaceuticals from the mucous membrane and, if the thickening polymer is used in combination with the cationic polymer, improve the retention in the mucous membrane by the action of the thickening polymer. Have been obtained, and the present invention has been completed. In addition, the present inventors have compared, among the cationic polymers, an aminoalkyl methacrylate copolymer or a polyvinyl acetate-l-ethylaminoacetate with a poly (L-arginine) which is also a cationic polymer. It has been found that the absorption promoting effect is excellent.

 That is, the present invention provides a powdery transmucosal administration preparation, particularly a nasal administration preparation, comprising a polymer drug and a cationic polymer. The powdery transmucosal administration preparation of the present invention may further contain a thickening polymer. Examples of the cationic polymer include aminoalkyl methacrylate copolymer, polyvinyl acetal getyl aminoacetate, and poly (L-arginine) aminoalkyl methacrylate copolymer. Preferred are vinyl and polyvinyl acetate acetylaminoacetates. Examples of the thickening polymer include hydroxypropyl methylcellulose. High molecular drugs can be selected from the group consisting of peptides and proteins having biological activity. Antibodies, vaccines, and antigens. The powdery transmucosal administration preparation of the present invention is particularly suitable for granulocyte colony stimulating factor. It is effective for transmucosal administration of insulin, erythropoietin, growth hormone or influenza antigens, especially for nasal administration.

Further, the present invention provides a powdery pharmaceutical composition comprising a high-molecular-weight drug and a cationic polymer. In the powdered pharmaceutical composition of the present invention, the high molecular drug is a peptide or protein having a physiological activity, an antibody, The powdered pharmaceutical composition of the present invention is particularly effective for administration of granulocyte colony stimulating factor, inulin, erythropoietin, growth hormone or influenza antigen, although it can be selected from the group consisting of vaccines and antigens. It is.

 Hereinafter, the present invention will be described in detail.

 In one embodiment of the present invention, the powdery transmucosal administration preparation of the present invention further comprises an excipient (for example, a saccharide) and a cationic polymer or a further thickening polymer in addition to a polymer drug. It can be obtained by freeze-drying (freeze-drying) or spray-drying (spray-drying) with the appropriate additives.

The polymeric drug used in the present invention refers to peptides and proteins having biological activity, antibodies, vaccines, antigens and the like, and specifically includes the following: It is not limited at all. Calcitonin, insulin, proinsulin, vasoplethsin, desmopressin, luteinizing hormone, luteinizing hormone-releasing hormone, somatostatin, prolactin, glucagon, gastrin, secretin, kallikrein, perokinase, neurotensin, enkepha. Lin, Kyoto torphin, Endorphin, Endothelin, Angiotensin, Transferrin, Atrial sodium diuretic peptide, Epidermal growth factor, Growth hormone, Parathyroid hormone, Interferon, Interleukin, Tumor Necrosis factor, leukemia cell inhibitory factor, blood stem cell proliferation factor, erythropoietin, granulocyte colony stimulating factor, granulocyte macrophage stimulating factor, macrophage colony stimulating factor, thrombopoetin Super Dose Dismutase, Tissue Plasminogen Activator, Antithrombin, Blood Coagulation Factor, Anti-IgE Antibody, Anti-IgA Antibody, Antitumor Antibody, Tumor Necrosis Factor Antibody, Anti-Interleukin Antibody, In HIV Japanese antibody, anti-platelet antibody, anti-hepatitis virus antibody, hepatitis vaccine, influenza vaccine (influenza antigen), pertussis vaccine, gif Peptides or proteins that can act as antigens and their hapten conjugates, such as terrier vaccines, tetanus vaccines, cedar pollen or poppy pollen, as well as mixtures of them with adjuvants. Further, it is easily presumed that the present invention can further increase the absorbability of the mucous membrane, particularly the nasal mucosa, even for pharmaceuticals having a smaller molecular weight than the above-mentioned high molecular weight pharmaceuticals. The application of the invention is considered useful.

 Examples of G-CSF, which is one of the high molecular drugs that can be used in the present invention, include a polypeptide having human G-CSF activity represented by the amino acid sequence of SEQ ID NOS: 1 to 3. Or a glycoprotein having a sugar chain attached thereto. Furthermore, a G—CSF derivative having G—CSF activity in which a part of the amino acid sequence having the same sequence is modified (substitution, deletion, insertion, and / or addition) is also included in G_CSF in the present invention.

 These G-CSFs can be extracted and separated from natural products, purified, or produced by transformants obtained by genetic transformation, and isolated and purified. . Examples of the host cell include Escherichia coli and mammalian cells (C127, CHO cells, etc.). Details of these production methods are described in, for example, JP-A-63-50066, JP-A-62-23964, JP-A-62-236. No. 488, and Japanese Patent Application Laid-Open No. Sho 63-26792.

The content of the high molecular drug in the powdery transmucosal administration preparation of the present invention is usually from 0.01 to 90 WZW%, preferably from 0.1 to 50 WZW%. The cationic polymer used in the present invention means a polymer having a cationic charge in a basic unit having a repeating structure or a structure having a cationic charge when dissolved. The cationic polymer used in the present invention may be any as long as it has an effect of promoting the absorption of the high molecular drug from the mucous membrane. Specifically, aminoalkyl methacrylate copolymer, polyvinyl acetal getyl amino acetoate, poly-L-arginine and the like can be used. Amino Archi The methacrylate copolymer can be obtained, for example, from Rohm Pharraa under the trade names Odragit E and Oudragit RS. Eudragit E is a copolymer of methyl methacrylate and butyl methacrylate and dimethylaminoethyl methacrylate, and has an average molecular weight of 150,000. Polyvinylacetate-l-ethylamino-acetate is, for example, a force available from Sankyo Co., Ltd. under the trade name of AEA. This is one of the acetal and hydroxyl groups obtained by dehydrating polyvinyl alcohol and acetoaldehyde. It is a polymer with an average molecular weight of 65,000, which can be obtained by ester-bonding getylaminoacetic acid to its part. Poly-L-arginine is a polymer of L-arginine and has an average molecular weight of 1000 to 1,000,000, preferably 12,100 to 92,000. More preferably, it is 92,000. Poly-L-arginine can be obtained from Sigma. The content of the thionionic polymer in the powdery transmucosal administration preparation of the present invention is usually 0.1 to 90%, preferably 1 to 50%.

 The thickening polymer used in the present invention refers to a polymer having viscosity when dissolved or swollen. The viscosity-enhancing polymer used in the present invention may be any one that increases the absorption of a high-molecular-weight drug when used in combination with a cationic polymer, compared to when the cationic polymer is used alone. Specifically, hydroxypropylmethylcellulose, hydroxypropylcellulose, carboxyvinyl polymer, agar powder, and arabia rubber powder can be used. The content of the thickening polymer in the powdery transmucosal administration preparation of the present invention is usually 0.1 to 90 WZW%, preferably 1 to 50 W / W%.

The excipient used in the present invention is typified by saccharides. Examples of such saccharides include xylitol, fructoses, sorbitol, lactose, inositol, sucrose, mannitol and the like. Can be Other excipients include starches, minerals, organic acids, and And the like. Starches include corn starch, wheat starch, potato starch, and the like. Inorganic substances include calcium phosphate, calcium hydrogen phosphate, sodium dihydrogen phosphate, sodium dihydrogen phosphate, magnesium carbonate, sodium chloride, sulfuric acid ruthenium, etc. It is. The organic acids include succinic acid, tartaric acid, citric acid, fumaric acid, lingic acid, dalconic acid, glucuronic acid and salts thereof. The amino acids include L-arginine, D, L-methionine, L-phenylalanine, L-glutamic acid, and the like. The content of the excipient in the powdery transmucosal administration preparation of the present invention is usually 1 to 90 W / W%, preferably 5 to 80 WZW%.

 In the present invention, additives such as lubricants are used as needed. Lubricants include magnesium stearate, stearate, talc, and the like. The content of the additive in the powdery transmucosal administration preparation of the present invention is usually 0.01 to 90 W / W%, preferably 0.05 to 50 W ZW%.

 Hereinafter, an example of the method for producing the powdery transmucosal administration preparation of the present invention will be briefly described.

 Mix the G-CSF buffer solution with a buffer solution in which a cationic polymer and excipients such as sucrose and mannitol or a thickening polymer have been dissolved. This is spray-dried to obtain a powder. A required amount of the obtained powder is weighed and filled into a capsule to obtain a powdery transmucosal administration preparation.

The powder of the preparation for transmucosal administration produced as described above usually has a particle size (diameter) of 0.1 to 500; am, and preferably has a particle size of 5 to 100 µm. The powder of the transmucosal administration formulation is easy to handle if it is made into a forcepsell. Examples of the capsule base material include gelatin, hydroxypropyl methylcellulose, methylcellulose, starch and the like.These materials include glycerin, sorbitol, carrageenan, polyethylene glycol, and polyethylene glycol. Gum arabic may be added to increase plasticity. In addition, potassium chloride, sucrose, coloring agents, and titanium oxide may be added.

 The powdery transmucosal administration preparation of the present invention can be administered to a patient's mucosa when necessary or at an appropriate administration frequency. Specific examples of mucous membrane include nasal mucosa, ocular mucosa, oral mucosa, lung mucosa, vaginal mucosa, and gastrointestinal mucosa such as gastric mucosa, small intestinal mucosa, large intestinal mucosa, and rectal mucosa. it can. For example, when the formulation of the present invention is to be administered nasally, a forceps containing the powdered formulation is set in a small nebulizer (publicizer), a hole is made in the capsule, and a nozzle is inserted into the nostril. By holding the rubber ball while breathing in with the nose, the powdery preparation may be sprayed into the nasal cavity. Formulations containing granulocyte colony stimulating factor as an active ingredient may be used 1 to 4 times a day, in an amount of 1 to 500 g / kg / day, preferably 5 to 100 tg / kg / day in terms of the amount of the active ingredient. It can be administered to patients. Formulations containing insulin as an active ingredient can be administered to patients at a dose of 0.1 to 100 U / kg / day, preferably 0.5 to 20 U / kg / day, 1 to 4 times a day. May be administered. Formulations containing erythropoietin as an active ingredient can be administered 1 to 4 times a day in an amount of 50 to 50,000 IU / kg / day, preferably 200 to 8,000 IU / kg / day, in terms of active ingredient. Then, I'll give it to the patient. Formulations containing growth hormone as an active ingredient may be administered 1 to 4 times daily, in an amount of 0.1 to 50 IU / kg / day, preferably 0.4 to 15 IU / kg / day, of active ingredient, It may be administered to a patient. Preparations containing influenza antigen as an active ingredient should be administered 1 to 4 times a day at intervals of 2 to 6 weeks, in terms of the amount of the active ingredient of 0.5 to 200 CCA / kg / day, preferably 2 to 4 A dose of 40 CCA / kg / day can be administered to the subject.

This description includes part or all of the contents as disclosed in the description and Z or drawings of Japanese Patent Application Nos. 10-192722 and 11-81549, which are the basis of the priority of the present application. BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described specifically with reference to Examples, but the scope of the present invention is not limited to these Examples.

 The cationic polymer, sucrose, D-mannitol, hydroxypropyl methylcellulose, sodium hyaluronate, and their buffer solution components (buffer components) used in the following Examples and Comparative Examples are as follows. .

Cationic polymer

 Poly — L-arginine (Sigma)

 Aminoalkyl methacrylate copolymer E (Rohm Pharma, trade name: Eudragit E100)

 Polyvinyl acetal getylamioacetate (Sankyo Co., Ltd., product name: AEA)

 Jetylaminoethyl (DEAE) -dextran (Fluka) Chitosan (Chitosan 8B, manufacturer: Katokichi, sales: Funakoshi)

)

Sucrose (Kosakai Pharmaceutical, Japanese Pharmacopoeia white sugar)

 D-Mannitol (Kao Nikoku Mannitol Kao-I) Hydroxypropylmethylcellulose (Shin-Etsu Chemical, Trade Name: Metro

60SH4000)

Sodium hyaluronate (Tokyo Chemical Industry)

Buffer component

 Cuic acid (Toyo Pharmaceutical), Phosphoric acid (Domestic Chemicals)

 The high molecular drugs used in the following examples are as follows.

Granulocyte colony stimulating factor (G-CSF) is a polypeptide having the amino acid sequence of SEQ ID NO: 1 produced by genetically modified Escherichia coli (see JP-T-63-500636). The G-CSF thus obtained was concentrated and replaced with a buffer to obtain a G-CSF buffer.

Insulin is commercially available (Human recombinant from Behringer Mannheim) : ^ = About 5700). Erythropoietin is a commercially available product (Kirin Brewery human recombinant Mw = about 30,000).

Growth hormone is commercially available (Chemicon human recombinant Mw = about 22,200).

Influenza A antigen is commercially available (Chemicon). Example 1>

 A buffer solution of sucrose and poly-L-arginine was added to a buffer solution of granulocyte colony-stimulating factor (G-CSF) and spray-dried to obtain a powdery nasal preparation having the following formulation.

 G-C S F 20 W / W%

 Poly — L—Arginine 20 W / W%

 Sucrose 26 W / W%

 Buffer component

 100 W / W% Example 2>

 To a buffer solution of granulocyte colony-stimulating factor (G-CSF) was added a buffer solution of sucrose and polyvinylacetate-l-jetilaminoacetate (AEA), and spray-dried to obtain a powdery nasal preparation having the following formulation .

 G-C S F 20 W / W%

 A E A 20 W / W%

 Sucrose 2 6 WZW%

 Buffer component appropriate amount

 100% w / w% Example 3>

Sucrose and granulocyte colony stimulating factor (G-CSF) buffer solution And aminoalkyl methacrylate copolymer E (Eudragit E1

) And spray-dried to obtain a powdery nasal preparation having the following formulation.

 G-C S F 20 W / W%

 Oy Dragit E100 20 W / W%

 Sucrose 26 W / W%

 Buffer component appropriate amount

 100 w / w% Example 4>

 Add a buffer solution of sucrose and aminoalkyl methacrylate copolymer E (Eudragit E100) to a buffer solution of granulocyte colony stimulating factor (G-CSF), spray-dry, and powder with the following formulation A nasal preparation was obtained.

 G-C S F 20 W / W%

 Oy Dragit E100 10 w / w%

 Sucrose 6 3 W / W%

 Buffer component Mm.

 100 W / W% Example 5>

 Sucrose and aminoalkyl methacrylate copolymer E (Eudragit E1) were added to a buffer solution of granulocyte colony stimulating factor (G-CSF).

) And spray-dried to obtain a powdery nasal preparation having the following formulation.

 G-C S F 20 W / W%

 Oy Dragit E100 20 W / W%

Sucrose 5 3 W / W% Buffer component

 Total 100 W / W

Example 6>

 Add a buffer solution of sucrose and aminoalkyl methacrylate copolymer E (Eudragit E100) to a buffer solution of granulocyte colony stimulating factor (G-CSF), spray-dry, and powder with the following formulation A nasal preparation was obtained.

 G-C S F 20 W / W%

 Oy Dragit E100 30 W / W%

 Sucrose 4 3 W / W%

 Buffer component Mm.

 1 0 0 w / w%

Example Ί>

 To a buffer solution of granulocyte colony stimulating factor (G-CSF), add a buffer solution of sucrose and aminoalkyl methacrylate copolymer (Eudragit E100), spray-dry, and spray-dry the following formulation. A powdery nasal preparation was obtained.

 G-C S F 20 W / W%

 Oy Dragit E100 5 7 W / W%

 Buffer component appropriate amount

 1 0 0 W / W%

Example 8>

Add a buffer solution of sucrose and aminoalkyl methacrylate copolymer E (Eudragit E100) to a buffer solution of granulocyte colony stimulating factor (G-CSF), spray-dry, and mix Powdered nasal formulation Profit

 Was

G-C S F 20 W / W

 Oy Dragit E100 20 W / W

 Sucrose 4 7 W / W

 Buffer component

 100 W / W Example 9>

 A buffer solution of sucrose and aminoalkyl methacrylate copolymer E (Eudragit E100) and a buffer solution of hydroxypropyl methylcellulose (HPMC) are added to a buffer solution of granulocyte colony stimulating factor (G-CSF). In addition, it was spray-dried to obtain a powdery nasal preparation having the following formulation.

 G—C S F 20 W / W%

 Oy Dragit E100 20 W / W%

 H P M C 10 W / W%

 Schloss 3 7 W / W%

 Buffer component appropriate amount

 Total amount 1 0 0 wzw%

<Example 10>

 To a buffer solution of granulocyte colony stimulating factor (G-CSF), add a buffer solution of sucrose and aminoalkyl methacrylate copolymer E (Eudragit E100) and hydroxypropyl methylcellulose (HPMC). It was spray-dried to obtain a powdery nasal preparation having the following formulation.

 G-C S F 20 W / W%

 Oy Dragit E100 20 W / W%

 H P M C 20 W / W%

Sucrose 2 7 W / W% Buffer component

 1 0 0 w / w

Comparative Example 1>

 A sucrose buffer solution was added to a granulocyte colony stimulating factor (G-CSF) buffer solution, and spray-dried to obtain a powdery nasal preparation having the following formulation.

 G-C S F 20 W / W%

 Schloss 4 6 W "W%

 Buffer component appropriate amount

 1 0 0 W / W% Comparative Example 2>

 A buffer solution of sucrose and sodium hyaluronate was added to a buffer solution of granulocyte colony stimulating factor (G-CSF), and spray-dried to obtain a powdery nasal preparation in the following manner.

 G-C S F 20 W / W%

 Sodium hyaluronate 20 W / W%

 Sucrose 26 W / W%

 Buffer component

 1 0 0 W / W%

<Experimental example 1>

Using male beagle dogs, the gelatin capsules were filled with the preparations prepared in Example 1 and Comparative Example 1 so as to have a G-CSF of 100 gg / kg. Gelatin capsules were attached to a publisher (R) (Ishikawa Seisakusho) with a 2.5 cm length of silicon tube adhered to the tip to prepare for administration. The silicon tube part of the publisher was inserted into the nasal cavity from the nostril, and the rubber ball was pressed to administer. Forearm vein at regular intervals after administration More blood was collected. 0 in the blood. The 5? Concentration was measured using the 1 SA method (T. Ichikawa et al .: Experimental Hematology 23: 192-195 (1955)). Table 1 shows the values of the area under the blood G—CSF concentration—time curve (AU C _G ). In this result, the addition of Example 1 shows a high value in AU C _G, poly one L- arginine as a cationic polymer compared to Comparative Example 1 containing no high molecular compound containing a poly _ L one-arginine G-CSF was found to be promoted through the nasal mucosa. table 1

 Dosage formulation Comparative Example 1 Example 1

AUC _G 0 → 32 hours 5.3 1 0.3

(-hr-mr ¹ ) Experiment 2>

Using male beagle dogs, the gelatin capsules were filled with the preparations prepared in Comparative Examples 1 and 2 so that the G-CSF became 100 g / kg. Gelatin capsules were attached to a bubbler (R) (manufactured by Ishikawa Seisakusho) with a silicon tube of about 2.5 cm length attached to the tip to prepare for administration. The silicone tube portion of the publisher was introduced into the nasal cavity through the nostrils, and the rubber ball portion was pressed for administration. After the administration, blood was collected from the forearm vein at regular intervals. G-CSF concentration in blood was measured by ELISA. Table 2 shows the values of the area under the blood G—CSF concentration—time curve (AU C _G ). As a result, Comparative Example 2 containing sodium hyaluronate, which is a polymer that is not a cationic polymer, showed almost the same value in AUC (; No effect was shown. Table 2

 Dosage formulation Comparative Example 1 Comparative Example 2

AUC _G 0 → 32 hours 4.8.4.6 (ng · hr -πιΓ ¹ )

Using a male beagle dog, the preparations prepared in Examples 1, 2 and 3 were filled into gelatin capsules so that the G-CSF became 10 Qng / kg. Gelatin capsules were attached to a publisher (R) (manufactured by Ishikawa Seisakusho) with a silicon tube approximately 5.0 cm in length adhered to the tip to prepare for administration. The silicone tubing of the publisher was inserted into the nasal cavity through the nostrils, and the rubber ball was pressed to administer. After the administration, blood was collected from the forearm vein at regular intervals. The number of leukocytes in the collected blood was counted using a microcell counter. G-CSF concentration in blood was measured using ELISA method. Table 3 shows the area under the increasing white blood cell count-time curve (AU _CW ) and the area under the G-CSF concentration-time curve in blood (AU CJ). As a result, it was found that AEA and Eudragit E100 have a better absorption promoting effect than poly-L-arginine.

 Dosage formulation Example 1 Example 2 Example 3

AUC _w 0 → 72 hours 3 0 8 3 5 0 9 5 5 7 0 7

(Count 'hr-ml- ¹ )

AUC _e 0 → 31 hours 1 3.4 7 3.0 44.4

(ng-hr-πιΓ ¹ ) Using male beagle dogs, the preparations prepared in Examples 4, 5, 6, and 7 were filled into gelatin capsules so as to have a G-CSF of 100 βg / kg. A gelatin capsule was attached to a publisher (R) (manufactured by Ishikawa Seisakusho) with a silicon tube of about 5.0 cm length adhered to the tip to prepare for administration. The silicon tube portion of the repurifier was inserted into the nasal cavity from the nostril, and administration was performed by pressing the rubber ball portion. After the administration, blood was collected from the forearm vein at regular intervals. The number of leukocytes in the collected blood was counted using a microcell counter. G-CSF concentration in blood was measured by ELISA. Table 4 shows the values of the area under the increased white blood cell count hourly curve (AUC _W ) and the area under the blood G_CSF concentration-time curve (AU C _G ). As a result, even when the content of Eudragit E100 was varied, the effect was maintained. Table 4

 Dosage formulation Example 4 Example 5 Example 6 Example Ί

AUC _W 0 → 72 hours 3 4 7 5 4 0 5 3 4 1 3 8 4 5 6 2 (count 'hr'ni ¹ )

AUCG 0 → 31 hours 2 5 .6 2 7 .1 1 6 .8 ^{1 1} .1 (ng-hr -ηιΓ ¹ )

<Experimental example 5>

Using male beagle dogs, the gelatin capsules were filled with the preparations prepared in Examples 8, 9 and 10 so as to have a G-CSF of 100 / ag / kg. A gelatin capsule was attached to a bubbler (R) (manufactured by Ishikawa Seisakusho) with a silicon tube of about 5.0 cm length adhered to the tip to prepare for administration. The silicone tube portion of the publisher was introduced into the nasal cavity through the nostrils, and the rubber ball portion was pressed for administration. After the administration, blood was collected from the forearm vein at regular intervals. Use a microcell counter to determine the number of white blood cells in the collected blood. And counted. G-CSF concentration in blood was measured by ELISA. Table 5 shows the values of the area under the increased white blood cell count-time curve (AU _CW ) and the area under the blood G-CSF concentration-one-hour curve (AU C _G ). As a result, it was clarified that the effect of adding HPMC together with Eudragit E100 was stronger than that of Eudragit alone. Table 5

 Dosage formulation Example 8 Example 9 Example 10

AUC _w 0 → 72 hours 3 9 8 8 5 4 8 2 5 6 1 8

(Count 'hr'ml—

AUC _G 0 → 31 hours 1 9.95 5 4.5 76.7 (ng · hr -ml " ¹ ) Example 1 1〉

 To a buffer solution of granulocyte colony stimulating factor (GCSF), add a buffer solution of D-mannitol and aminoalkyl methacrylate copolymer E (Eudragit E100), spray-dry, and administer powdered nasally of the following formulation A formulation was obtained.

 G-C S F 10.0 W / W%

 Oil Drag E 1 0 0 7.5 W / W%

 D—mannitol 75.0 W / W%

 Buffer component

 1 0 0 W / W%

<Comparative Example 3>

A buffer solution of D-mannitol was added to a buffer solution of granulocyte colony stimulating factor (G-CSF) and spray-dried to obtain a powdery nasal administration preparation having the following formulation. G-CSF 10.0 W / WD — Mannitol 81.8 W / W

 Buffer component

 100 W / W%

<Experimental example 6>

 The preparations prepared in Example 11 and Comparative Example 3 were filled into gelatin capsules using male beagle dogs so that 50 ng / kg of G-CSF was administered. Gelatin capsules were attached to a publisher (R) (manufactured by Ishikawa Seisakusho) with a silicon tube approximately 5.0 cm in length adhered to the tip to prepare for administration. The silicon tube part of the publisher was inserted into the nasal cavity from the nostril, and the rubber ball was pressed to administer. After the administration, blood was collected from the forearm vein at regular intervals. Blood G-CSF concentration was measured using the ELISA method. Table 6 shows the values of the area under the blood G—CSF concentration one-hour curve (AUCG). As a result, it was found that Eudragit E100 significantly promoted the nasal absorption of G-CSF. Table 6

 Dosage form Example 1 1 Comparative example 3

AUC _G 0 → 31 hours 6 7 .1 1 6 4

(ng-hr-ml ^-1 ) Example 1 2>

 A buffer solution of sucrose and aminoalkyl methacrylate copolymer E (Eudragit E100) and hydroxypropyl methylcellulose (HPMC) is added to a buffer solution of Insulin, and the mixture is spray-dried. A nasal formulation was obtained.

Insulin 18 W W% Oil Drag E 1 0 0 2 7 W / W

 H P M C 9 W / W

 Sucrose 3 2 W / W (buffer component

 100 W / W Example 1 3>

 A buffer solution of sucrose, poly-L-arginine and hydroxypropylmethylcellulose (HPMC) was added to the buffer solution of Insulin, and the mixture was spray-dried to obtain a powdery nasal preparation having the following formulation.

 Insulin 18 W W%

 Poly — L arginine 2 7 W Z W%

 H PMC 9 W / W%

 Sucrose 3 2 WZW%

 Buffer component appropriate amount

 Total amount 100 W / W% Example 1 4>

 Add sucrose and dexamylaminoethyl (DEAE) -dextran and hydroxypropyl methylcellulose buffer (HPMC) buffer solution to Insulin buffer solution, spray-dry, and powder nasal A dosage formulation was obtained.

 Insulin 18 W / W%

 D E A E—Dextran 2 7 W / W%

 H PMC 9 W / W%

 Sucrose 3 2 W / W%

 Buffer component

1 0 0 w / w% Example 1 5>

 A buffer solution of sucrose, chitosan, and hydroxypropylmethylcellulose (HPMC) was added to the buffer solution of insulin, and spray-dried to obtain a powdery nasal preparation having the following formulation.

 Insulin 18 W / W%

 Chitosan 27 W / W%

 H PMC 9 W / W%

 Sucrose 3 2WZW%

 Buffer component appropriate amount

 100 W / W% Comparative Example 4>

 A buffer solution of sucrose and hydroxypropylmethylcellulose (HPMC) was added to the buffer solution of insulin, and the mixture was spray-dried to obtain a powdery intranasal preparation in the following manner.

 Insulin 18 W W%

 H PMC 9 W / W%

 Sucrose 60 WZWO / O Buffer component

 100% w / w% Comparative Example 5>

 Inulin was dissolved in a buffer solution to prepare a subcutaneous administration solution having the following concentrations.

Insulin 1.0 mg / m 1 Experimental Example 7> Using the male beagle dog, the preparations prepared in Examples 12, 13, 13, 14, and 15 and Comparative Example 4 were intranasally administered, and the preparation prepared in Comparative Example 5 was subcutaneously administered. The nasal administration group was filled into gelatin capsules so that 70 μg / kg of insulin was administered. Gelatin capsules were attached to a publisher (R) (manufactured by Ishikawa Seisakusho) with a silicon tube of about 5.0 cm length adhered to the tip to prepare for administration. The silicone tube part of the publisher was inserted into the nasal cavity from the nostril, and the rubber ball was pressed to administer the drug. In the subcutaneous administration group, the preparation of Comparative Example 5 was subcutaneously administered as insulin to the back of a 25 μg / kg bi-dog dog. After administration, blood was collected from the forearm vein at regular intervals. Blood insulin concentration was measured using the ELISA method. Table 7 shows the values of the area under the blood insulin concentration one-hour curve (AUC). As a result, it was found that Eudragit E100 significantly promoted the nasal absorption of Insulin. Furthermore, its absorption promoting effect was superior to that of poly-L-arginine, DEAE-dextran and chitosan. The bioavailability of the preparation of Example 12 for subcutaneous administration was 27%. Table 7

 Dosage formulation Example 1 2 Comparative example 4 Comparative example 5

AUC 0 → 7 hours 29.0 0 2.6 38.8 (ng-hr-ml " ¹ ) Dosage form Example 1 3 Example 1 4 Example 1 5

AUC 0 → 7 hours 2.9 2.5.2 23.6 (ng'hr 'ml- ¹ )

Granulocyte colony stimulating factor (G-CSF) in buffer solution contains D-mannitol and aminoalkyl methacrylate copolymer E (Dragon E 100) was spray-dried to obtain a powdery nasal administration formulation having the following formulation.

G-C S F 10.0 W / W Eudragit E 1 0 0 7.5 W / W D-mannitol 75.2 W / W Buffer component

 1 0 0.0 W / W Example 1 7>

 A buffer solution of D-mannitol and poly-L-arginine was added to a buffer solution of granulocyte colony stimulating factor (G-CSF), and the mixture was spray-dried to obtain a powdery nasal administration preparation having the following formulation.

 G-C S F 10.0 w / w% Poly L-arginine 7.5 W / W%

D-mannitol 7 5.2 W / W% buffer component

 1 0 0.0 w / w%

<Example 18>

 A buffer solution of granulocyte colony stimulating factor (G-CSF) was added to a buffer solution of D-mannitol and getylaminoethyl (DEAE) -dextran, and the mixture was spray-dried to obtain a powdery nasal preparation having the following formulation. .

 G-C S F 10.0 W / W%

D E A E—dextran 7.5 W / W% D—mannitol 75.2 W / W% Buffer component

Whole amount 1 0 0.0 WZW% <Example 19>

 A buffer solution of D_mannitol and chitosan was added to a buffer solution of granulocyte colony-stimulating factor (G-CSF), and the mixture was spray-dried to obtain a powdery nasal administration preparation having the following formulation.

 G -C S F 10.0 W / W% Chitosan 75 W / W% D — Mannitol 752 W / W% Buffer component

 10.0 0.0 W / W% Experimental Example 8>

The preparations prepared in Examples 16, 17, 18, and 19 using male beagle dogs were filled into gelatin capsules so as to be administered at 50 g / kg as G-CSF. Gelatin capsules were attached to a publisher (R) (manufactured by Ishikawa Seisakusho) with a silicon tuple of about 5.0 cm length attached to the tip to prepare for administration. The silicon tube of the republizer was inserted into the nasal cavity from the nostrils and the rubber ball was pressed to administer the drug. After the administration, blood was collected from the forearm vein at regular intervals. G-CSF concentration in blood was measured using ELISA method. Table 8 shows the values of the area under the blood G-CSF concentration hour curve (AUCG). As a result, it was found that the powder under Eudragit E 100 had the highest area under the one-hour curve of blood G—CSF concentration (AUC _E ). From the above, it was clarified that the absorption promoting effect of Eudragit E100 was superior to that of other polycations such as poly-L-arginine, DEAE-dextran and chitosan. Table 8

 Dosage formulation Example 16 Example 17

AUC _G 0 → 31 hours 4 6. 4 3 5. 7

(ng · hr · ml " ¹ )

Dosage form Example 18 Example 19

AUC _G 0 → 31 hours 2 2.9 4 2.8

ng · hr · ml ” ¹ ) Example 20>

 To a buffer solution of erythropoietin was added a buffer solution of sucrose and aminoalkyl methacrylate copolymer E (Eudragit E100) and hydroxypropyl methylcellulose (HPMC), and the mixture was spray-dried. Was obtained in the form of a powder.

 Erythropoietin 3 0 W / W%

 Oil Drag E 1 0 0 3 0 W / W%

 H P M C 10 w / w%

 Sucrose 15 W / W%

 Buffer component

 1 0 0 w / w% Comparative Example 6>

 A buffer solution of sucrose and hydroxypropylmethylcellulose (HPMC) was added to a buffer solution of erythropoietin, and the mixture was spray-dried to obtain a powdery intranasal preparation having the following formulation.

Erythropoietin 30 W / W% HPMC 10 W / W

 Sucrose 4 5 W / W

 Buffer component

 1 0 0 W / W

<Example 21>

 A buffer solution of sucrose, poly-L-arginine, and hydroxypropylmethylcellulose (HPMC) was added to a buffer solution of Eris's mouth poetin, and spray-dried to obtain a powdery nasal preparation having the following formulation.

 Erythropoietin 3 0WZW%

 Poly — L—Arginine 3 0 W Z W%

 H P M C 10 W / W%

 Sucrose 15 W W%

 Buffer component appropriate amount

 1 0 0 W / W%

<Example 22>

 To a buffer solution of erythropoietin, add a buffer solution of sucrose and getylaminoethyl (DEAE) -dextran and hydroxypropylmethylcellulose (HPMC), spray-dry, and powder nasal administration of the following formulation A formulation was obtained.

 Erythropoietin 30 W / W%

 D E AE—Dextran 30 W / W%

 H P M C 10 W / W%

 Sucrose 15 W / W%

 Buffer component

1 0 0 w / w% <Example 23>

 A buffer solution of sucrose, chitosan, and hydroxypropylmethylcellulose (HPMC) was added to a buffer solution of erythropoietin, and the mixture was spray-dried to obtain a powdery nasal preparation having the following formulation.

Erythropoietin 3 OWZW ⁰ /)

 Chitosan 30 W / W%

 H P M C 10 W / W%

 Schloss 1 5 W / W%

 Buffer component appropriate amount

 Total volume 100 W / W%

<Comparative Example 7>

 Erythropoietin was dissolved in a buffer solution to prepare a subcutaneous administration solution having the following concentration.

 Erythropoietin 2 5 0 μ 8 / πι 1

<Experimental example 9>

Using the male beagle dog, the preparations prepared in Examples 20, 21, 22, 23 and Comparative Example 6 were intranasally administered, and the preparation prepared in Comparative Example 7 was subcutaneously administered. For the nasal administration group, gelatin capsules were filled so that 1 SO ^ gZ kg as erythropoietin was administered. Gelatin capsules were attached to a publisher (R) (manufactured by Ishikawa Seisakusho) with a silicon tube of about 5. O cm length adhered to the tip to prepare for administration. The silicone tube part of the publisher was inserted into the nasal cavity from the nostril, and the rubber ball was pressed to administer the drug. In the subcutaneous administration group, the preparation of Comparative Example 7 was subcutaneously administered as erythropoietin to the back of 5 μg Z kg beagle dog. After the administration, blood was collected from the forearm vein at regular intervals. The blood erythropoietin concentration was measured using an ELISA method. Table 9 shows the values of the area under the erythropoietin concentration-hour curve (AU C) in blood. did. As a result, it was found that Eudragit E100 significantly promotes nasal absorption of erythropoietin. Furthermore, its absorption promoting effect was superior to poly-L-arginine, DEAE-dextran and chitosan. The bioavailability of Example 20 for subcutaneous administration was 15%. Table 9

 Dosage formulation Example 20 Comparative Example 6 Example 21

 A U C 0 → 11 hours 2 9 1 1.5 19.1

(U-hr-ml " ¹ )

Dosage formulation Example 2 2 Example 2 3 Comparative example 7

 AU C 0 → 11 hours 6.6.2 0.2 7.9

(U-hr-ml " ¹ )

(Erythropoietin 1 _ag is equivalent to 130 U) Example 24>

 D_mannitol and amino in buffer solution of growth hormone

A buffer solution of acrylate copolymer E (Eudragit E100) was added and spray-dried to obtain a powdery nasal administration formulation having the following formulation.

 Growth hormone 10.0 WZW% Oy dragit E 1 0 0 7.5 W / W%

D-mannitol 7 5.2 W / W% buffer component

1 0 0.0 W / W% Comparative Example 8> A buffer solution of D-mannitol was added to a buffer solution of growth hormone, and the mixture was sprayed and dried to obtain a powdery intranasal preparation having the following formulation.

 Growth hormone 1 0.owzw%

D-mannitol 8 2.7 W / W% buffer component

 1 0 0.0 w / w% Experimental example 1 0>

 Using male beagle dogs, the gelatin capsules were filled with the preparations prepared in Example 24 and Comparative Example 8 so that 50 zg Zkg was administered as a growth hormone. A gelatin capsule was attached to a publisher (R) (manufactured by Ishikawa Seisakusho) with a silicon tube of about 5.0 cm in length attached to the tip to prepare for administration. The silicone tubing of the publisher was inserted into the nasal cavity through the nostrils and the rubber ball was pressed to administer the drug. After the administration, blood was collected from the forearm vein at regular intervals. Blood growth hormone levels were measured using the ELISA method. Table 10 shows the values of the area under the blood growth hormone concentration one hour curve (AU C). As a result, it was found that Eudragit E100 significantly promoted the nasal absorption of growth hormone, and the absorption rate was a very high value of 10 times that of the case without the addition. Table 10

 Dosage formulation Example 2 4 Comparative example 8

 AU C 0 → 11 hours 1 3 0 1 3

(, ng-hr -ml " ¹ ) Example 25>

Buffer D-mannitol and aminoalkyl methacrylate copolymer E (Eudragit E 100) in buffer solution of influenza A antigen. The resultant was dried by drying to obtain a powdered nasal preparation having the following formulation. However, the weight of influenza A antigen in the following table is the value including the buffer component in the reagent.

 Influenza A antigen 4.0 W / W% Eudragit E 1 0 0 7.5 W / W%

D-mannitol 8 1.2 W / W% buffer component weekly weight

 1 0 0.0 w / w%

<Comparative Example 9>

 A buffer solution of D-mannitol was added to a buffer solution of influenza A antigen, and the mixture was spray-dried to obtain a powdery nasal preparation having the following formulation. However, the weight% of influenza A antigen in the table below is a value including the buffer component in the reagent.

 Influenza A antigen 4.0 W / W%

D-mannitol 8 8.7 W / W% buffer component

 1 0.0 0.0 w / w% Example 2 6>

 A buffer solution of D-mannitol and poly-L-arginine was added to a buffer solution of influenza A antigen, and the mixture was spray-dried to obtain a powdery nasal administration formulation having the following formulation. However, the weight percentage of influenza A antigen in the table below is the value including the buffer component in the reagent.

 Influenza A antigen 4.0 W / W% Poly-L-arginine 7.5 WZ W%

D-mannitol 8 1.2 W / W% buffer component All: 1 0 0. 0 wzw%

<Example 27>

 A buffer solution of D-mannitol and getyl aminoethyl (DAE) -dextran was added to a buffer solution of influenza A antigen, and the mixture was spray-dried to obtain a powdery nasal administration formulation having the following formulation. However, the weight percentage of influenza A antigen in the table below is the value including the buffer component in the reagent.

 Influenza A antigen 4.0 W / W%

D E A E—dextran 7.5 W / W%

D-mannitol 8 1.2 W / W% buffer component appropriate amount

 Example 2 8> 10.0 0.0 W / W%

 A buffer solution of D-mannitol and chitosan was added to a buffer solution of influenza A antigen, and the mixture was spray-dried to obtain a powdery nasal administration preparation having the following formulation. However, the weight percentage of influenza A antigen in the table below is the value including the buffer component in the reagent.

 Influenza A antigen 4.0 W / W% Chitosan 7.5 W / W%

D-mannitol 8 1.2 W / W% buffer component appropriate amount

 1 0 0.0 W / W% Experimental example 1 1>

 << Day 1 of experiment (1st dose) >>>

 Blood was collected from the forearm vein of a male beagle dog used in the experiment. Example

25, 26, 27, 28, and Comparative Example 9 Gelatin capsules were filled in such a way that 24 μl equivalent of the A-antigen was administered. A gelatin capsule was attached to a bubbler (R) (manufactured by Ishikawa Seisakusho) with a silicon tube of about 5.0 cm length adhered to the tip to prepare for administration. The silicone tube portion of the publisher was introduced into the nasal cavity through the nostril, and the rubber ball was pressed to administer the drug.

<<Experiment 15 Day 5 (2nd administration) >>>

 The preparations prepared in Examples 25, 26, 27, 28 and Comparative Example 9 were administered nasally. In this case, the grouping, dosage, and administration method are the same as on the first day of the experiment.

<Day 9 of Experiment 2>

 Blood was collected from the forearm vein of male beagle dogs to which influenza A antigen was administered.

<<Measurement of antibody amount>>

On days 1 and 29, the amount of anti-influenza A antibody in the serum collected was measured by ELISA. The types of antibodies measured were IgG1 and IgG2, and changes in the amount of anti-influenza A antibody on day 1 were compared. The amount of the anti-influenza A antibody was compared with the absorbance difference between the well on which the antigen was immobilized and the well on which the antigen was not immobilized. The percentage of individuals in which anti-influenza A antibody was induced on Day 29 of the experiment is shown in Tables 11 and 12 (four animals in each group). As a result, in the Eudragit E100-added group, it was found that both anti-influenza A-IgG1 and IgG2 were most frequently induced. Thus, Eudragit E100 is useful as a nasal vaccine adjuvant, and its action is superior to other polycations such as poly-L-arginine, DEAE-dextran and chitosan. It became clear. Table 11: Anti-influenza A—IgG1 antibody induction rate

 Dosage formulation Example 25 Comparative Example 9 Example 26

2 9th day (after 2 sensitizations) 50% 0% 25%

Dosage formulation Example 2 7 Example 2 8

 2 Day 9 (after 2 sensitizations) 0% 50%

Table 12: Anti-influenza A—IgG2 antibody induction rate

 Dosage formulation Example 25 Comparative Example 9 Example 26

2 9th day (after 2 sensitizations) 100% 25% 100%

Dosage formulation Example 2 7 Example 2 8

 2 9th day (after 2 sensitizations) 25% 25%

Industrial applicability

 Addition of a cationic polymer (especially aminoalkyl methacrylate copolymer or polyvinyl acetate acetylaminoacetate) to a high-molecular drug or addition of a thickening polymer to a powdered drug By doing so, it was possible to effectively absorb the high molecular drug from the mucous membrane. All publications, patents, and patent applications cited herein are hereby incorporated by reference in their entirety.

Claims

 The scope of the claims I. A powdery transmucosal administration preparation comprising a polymer drug and a cationic polymer.  2. The powdery transmucosal administration preparation according to claim 1, further comprising a thickening polymer.  3. The powdery transmucosal administration preparation according to claim 1, wherein the cationic polymer is an aminoalkyl methacrylate copolymer.  2. The powdery transmucosal administration preparation according to claim 1, wherein the cationic polymer is polyvinyl acetate-l-ethylamyl acetate.  5. The powdery transmucosal administration preparation according to claim 1, wherein the cationic polymer is poly L-arginine.  6. The powdery transmucosal administration preparation according to claim 2, wherein the thickening polymer is hydroxypropylmethylcellulose.  7. The powdery transmucosal administration preparation according to claim 1, wherein the polymer drug is selected from the group consisting of peptides and proteins having biological activity, antibodies, vaccines, and antigens.  8. The powdery transmucosal administration preparation according to claim 7, wherein the peptide having physiological activity is a granulocyte colony-stimulating factor.  9. The powdery transmucosal administration preparation according to any one of claims 1 to 8, which is a nasal administration preparation.  10. A powdery pharmaceutical composition comprising a high molecular drug and a cationic polymer.  I I. The powdered pharmaceutical composition according to claim 10, wherein the polymer drug is selected from the group consisting of peptides and proteins having biological activity, antibodies, vaccines, and antigens. 12. The method of claim 11, wherein the polymeric drug is selected from the group consisting of insulin, erythropoietin, growth hormone and influenza antigen. Powdered pharmaceutical composition